title: Effects of in vivo exposure to bis(tri-n-butyltin)oxide, hexachlorobenzene, and benzo(a)pyrene on cytokine (receptor) mRNA levels in cultured rat splenocytes and on IL-2 receptor protein levels.
abstract: Analysis of cytokine (receptor) mRNA levels has been suggested to be a sensitive technique for predicting the immunomodulatory potential of drugs and chemicals. Furthermore, this type of analysis is thought to be important in unraveling mechanisms of immunotoxicity. To study these issues, male Wistar rats were exposed to the immunotoxic environmental contaminants bis(tri-n-butyltin) oxide (TBTO; 5, 20, or 80 mg/kg diet for 6 weeks), hexachlorobenzene (HCB; 50, 150, or 450 mg/kg diet for 6 weeks), or benzo(a)pyrene (B(a)P; 3, 10, 30, or 90 mg/kg body wt for 5 weeks by a daily (5 times a week) oral intubation). Spleen cells were cultured with Con A and analyzed by dot blot hybridization for IL-2, IFN-gamma, IL-2 receptor alpha-chain (IL-2R alpha; CD25), and IL-4 mRNA levels. In addition, spleen and thymus sections of TBTO-exposed animals were assayed immunohistochemically for CD25 expression. Exposure to TBTO resulted in a dose-dependent decrease in IL-2R alpha mRNA levels from 5 mg/kg, a dose-dependent increase in IFN-gamma mRNA levels from 20 mg/kg, and increased IL-2 mRNA levels at 80 mg/kg diet. Exposure to HCB resulted in a dose-dependent increase in IL-2 and IFN-gamma mRNA levels from 150 mg/kg and increased IL-2R gamma mRNA levels at 450 mg/kg diet. Exposure to B(a)P resulted in a dose-dependent increase in IL-2 and IFN-gamma mRNA levels from 10 mg/kg and increased IL-2R alpha mRNA levels at 90 mg/kg body wt. No effects were seen on IL-4 mRNA levels. Spleen and thymus sections of TBTO-exposed animals showed reduced CD25 expression from 5 mg/kg diet. These results show that (1) the correlation between altered cytokine (receptor) mRNA levels and functional endpoints is variable, depending on the type of functional endpoint tested and the compound studied, (2) these assays are among the most sensitive ones for TBTO and HCB immunotoxicity, and among the more sensitive ones for B(a)P immunotoxicity, and (3) for TBTO, these assays provide a possible clue to a mechanism for thymus atrophy, resulting from exposure to this compound: reduced IL-2R expression may impede thymocyte maturation, resulting in thymus atrophy.
authors.affiliation: Laboratory for Pathology and Immunobiology, National Institute of Public Health and the Environment, Bilthoven, The Netherlands. r.vandebriel@rivm.nl
authors.forename: R J
authors.initials: RJ
authors.lastname: Vandebriel
authors.forename: C
authors.initials: C
authors.lastname: Meredith
authors.forename: M P
authors.initials: MP
authors.lastname: Scott
authors.forename: P J
authors.initials: PJ
authors.lastname: Roholl
authors.forename: H
authors.initials: H
authors.lastname: Van Loveren
mesh_terms: D000818:Animals; D001564:Benzo(a)pyrene / Q000494:pharmacology*; D002273:Carcinogens / Q000494:pharmacology*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D005659:Fungicides, Industrial / Q000494:pharmacology*; D006581:Hexachlorobenzene / Q000494:pharmacology; D007150:Immunohistochemistry; D007376:Interleukin-2 / Q000378:metabolism*; D008297:Male; D009693:Nucleic Acid Hybridization; D009929:Organ Size / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D018121:Receptors, Cytokine / Q000235:genetics / Q000378:metabolism*; D015375:Receptors, Interleukin-2 / Q000378:metabolism; D013154:Spleen / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D013950:Thymus Gland / Q000378:metabolism / Q000473:pathology; D014220:Trialkyltin Compounds / Q000494:pharmacology
chemical_list: D002273:Carcinogens; D005659:Fungicides, Industrial; D007376:Interleukin-2; D012333:RNA, Messenger; D018121:Receptors, Cytokine; D015375:Receptors, Interleukin-2; D014220:Trialkyltin Compounds; C005961:bis(tri-n-butyltin)oxide; D001564:Benzo(a)pyrene; D006581:Hexachlorobenzene